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为进一步分析转AgGlpF基因耐盐转基因大豆E8A7027转化体的分子特征信息并对其进行特异性检测,本研究分离该转化体的旁侧序列并建立其特异性PCR检测体系。基于基因组重测序技术并结合Sanger测序技术,将基因组重测序分析获得的序列信息与参考大豆基因组进行对比,确定E8A7027转化体的T-DNA区在大豆基因组上的插入位点及其5′端和3′端的旁侧序列。根据旁侧序列设计PCR检测引物,并对检测体系的特异性、灵敏度进行实验室内验证。结果显示:转基因大豆E8A7027的整合位点为Chr09染色体的33886331位点,整合方式为单拷贝插入,受体基因组DNA在插入位点缺失了1段58 bp序列,分别获得E8A7027大豆的5′端旁侧序列910 bp和3′端旁侧序列802 bp。根据这两个旁侧序列设计引物建立的PCR检测方法,能够特异性地将E8A7027大豆转化体与其他转基因作物和非转基因大豆区分开,方法的检测灵敏度为0.05%。研究结果可为E8A7027转化体的安全评价和监管检测提供技术支撑。 相似文献
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转基因作物外源基因的表达是否会对土壤微生物生态系统产生不利影响是转基因作物环境安全评价中的重要一环,也关系到作物新品种的应用与推广.为此,本文采用第三代16S扩增子高通量测序技术,对转mEP-SPS基因的甘蓝型油菜甲HX6及其亲本甲9707不同发育阶段的根际土壤细菌群落进行了比较分析.结果显示,样品中油菜根际的优势细菌... 相似文献
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转基因作物安全评价是当前生物安全研究的重要内容。本实验以海南三亚南繁基地为实验平台,通过比较转BT/CPTI双价基因抗虫棉花SGK321和非转基因棉花石远321根际土壤微生物动态变化,初步研究了转基因棉花SGK321对农田生态环境的影响。通过常规培养计数、分子鉴定等手段监测土壤微生物多样性变化,经过近两年的跟踪研究,发现两次生长周期内,转基因棉花SGK321和非转基因棉花石远321根际土壤微生物(包括细菌、真菌、放线菌)的动态变化趋势大致相同,群落数量和种属组成都没有明显差异。此外,南繁大田种植环境条件下,除棉铃虫数量差异明显外,其余各种棉田虫害数量差异不明显。据此,我们初步推断南繁条件下转基因棉花SGK321的种植对土壤微生物及棉田虫害影响不显著。本研究可为在我国南繁基地开展转基因作物安全评价提供参考。 相似文献
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《中国油料作物学报》2021,(1)
发展转基因技术,占领转基因技术制高点,对增强我国农业国际竞争力具有重要意义;转基因技术及相关农产品国际贸易的快速发展,对转基因生物安全评估及监管提出了更高的要求。为此,编辑部特别邀请吴刚研究员组织本期专刊,旨在为我国转基因植物检测与安全评价提供更好的理论与技术支撑。主要内容包括新型转基因精准定量技术、标准物质研制、现场快速检测研究、分子特征验证等方面的研究论文和综述。 相似文献
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《大豆科学》2020,(1)
转基因作物的T-DNA插入及其与宿主基因组的连接序列构成的侧翼区域,是转基因事件检测的关键组成部分,也是转基因作物开发、评估和监管所必需的。前期研究得到2个携带大豆花叶病毒P3(soybean mosaic virus P3)基因干扰片段的转基因大豆事件,即L13和L104,可显著提高大豆对大豆花叶病毒、大豆花叶坏死病毒和西瓜花叶病毒的抗性,具有较强的光谱性。为了便于对该转基因事件进行监管以及建立事件特异性检测方法,本研究通过Illumina Xten平台,对L13和L104进行全基因组测序(whole genome sequencing,WGS),并结合生物信息学分析和PCR技术,对转基因事件L13和L104的旁侧序列进行了分析。通过全基因组测序,每一个事件获得了超过12.13 Gb的原始数据,测序深度为11×,与大豆参考基因组(Wm82.a2.v1)和转化载体序列做比较,初步识别出这两个转化事件T-DNA的边界及其旁侧序列。转化事件L13和L104的T-DNA分别位于Chr04和Chr11染色体上。进一步的PCR检测和序列测定表明转基因事件L13的T-DNA为单位点双拷贝整合到大豆基因组Chr04:46747946位点;转基因事件L104的T-DNA为单拷贝插入,整合位点为Chr11:30461895位点。结果证明了全基因组测序在识别转基因作物T-DNA插入和旁侧序列区域方面的有效性和稳定性。此外,插入位点的鉴定和事件特异性检测的建立将有助于广谱抗病毒转基因大豆品种的应用和发展。 相似文献
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转基因作物筛查用阳性质粒分子的构建及应用研究 总被引:1,自引:0,他引:1
通过分析全球商业化种植的转基因作物分子特征,选取常用的调控元件启动子CaMV35S、FMV35S和nos,终止子NOS、T-35S、g73′和E93′,标记基因bar、NPTII、hptII、pmi作为筛查靶标序列,通过重组PCR技术将这些元件克隆到双元载体pBI121上,构建成包含11个转基因元件的通用阳性质粒分子pBI121-ELEMENTS。质粒序列信息已提交至GenBank并获得序列号HM047294。该质粒分子对目前国内外批准的5类主要转基因作物(大豆、油菜、玉米、棉花和水稻)理论筛查覆盖率达到91.5%,对我国批准进口的27种转基因作物的理论筛查覆盖率达100%。以pBI121-ELEMENTS为阳性对照,筛查7个转基因品系,结果显示该分子能够有效用于转基因作物定性筛查。 相似文献
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Holden Verdeprado Tobias Kretzschmar Hasina Begum Chitra Raghavan Priya Joyce Prakash Lakshmanan 《Plant Production Science》2018,21(3):159-176
In the last decade, association mapping (AM) has become a well-established method to detect genes and quantitative trait loci (QTLs) associated with agronomically important traits. The identification of a large number of single nucleotide polymorphisms (SNPs) from genome sequencing and concurrent development of high-throughput genotyping platforms has led to AM being widely used for a range of crops. These technologies have been used in rice (Oryza sativa) to explore the abundant diversity and there is enormous potential to identify novel QTLs for traits of interest. Due to the availability of cost-effective high-throughput SNP genotyping methods and rapid developments in rice genomics, it is inevitable that these AM approaches will become more popular in the future, especially in the context of genome-wide association studies (GWASs). In this paper, we review the fundamental concepts, critical considerations and limitations of AM focusing on rice, and reiterate the importance of accurate phenotypic data. We also include a section about connecting GWAS to molecular breeding, covering practical consideration for breeders, which is required to use GWAS results in actual rice molecular breeding programs and which has not received adequate attention in the scientific literature. 相似文献
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遗传图谱构建与QTL定位对花生分子育种具有十分重要的意义.随着分子标记与测序技术的快速发展,极大地促进了花生重要数量性状的研究进程,并获得了一些与抗病、产量等重要性状相关联的分子标记.本文对国内外花生的分子标记开发、遗传图谱构建以及QTL定位的研究进展进行了综述,为花生分子标记辅助选择,提高花生育种效率,加速育种进程提供了理论参考. 相似文献
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《Journal of Crop Improvement》2013,27(1-2):213-226
Abstract A number of molecular marker techniques have been applied to improve the quality of cultivated lentil, aiding in the breeding and release of better adapted varieties and in quality assurance of export product. Continued increase in lentil production is dependent on the identification of present and potential limiting factors to the progression of lentil breeding. In particular, this chapter is concerned with limitations related to the threat from disease, limitations on the availability of genetic resources and quality assurance issues. The application of molecular markers in the development of strategies to help combat such threats is discussed. 相似文献
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Further improvement of rice productivity remains a challenge. Breeding is perceived as an important option to increase rice yield. However, the genetic progress of grain yield in most rice breeding programs was slow in the last decades. Although great progress in rice genomics and molecular biology has been achieved, the effect of such technological innovations on rice breeding is far small. Marker-assisted selection (MAS) for a few target quantitative trait loci (QTLs) has significant effects in improving qualitative traits, such as disease resistance. The success of MAS has therefore motivated breeders to identify and use major QTLs for yield and yield component traits. In this review, we summarized the recent methods in QTL identification, including novel statistical methods for linkage and association mapping, special population types, and whole-genome sequencing. We reviewed the successful application of marker-assisted gene introgression and gene pyramiding to improve grain yield and discussed the design of efficient MAS schemes to further increase the success rate of breeding programs. The use of well-characterized major QTLs through introgression and gene pyramiding is proven effective in improving grain yield, particularly yield under abiotic stress. Major QTLs that are stable across genetic background and growing environments are often found in less adapted germplasms, such as landraces and wild relatives. Advanced backcross QTL analysis and introgression lines, which integrate QTL discovery and utilization, are important methods for exploiting major QTLs contained in such germplasms. Next-generation sequencing substantially increases mapping resolution and accelerates the identification of casual genes underlying major QTLs. Practical guidelines derived from theoretical and empirical studies are given to guide the design of efficient marker-assisted gene introgression and pyramiding schemes. 相似文献
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基因型依赖及白苗再生等问题限制了花药离体培养技术潜力的充分发挥。为了克服这些问题,提高花药离体培养效率,国内外学者采用传统遗传学、细胞学、生理生化、分子标记、基因克隆、转录组学及蛋白组学等方法,在小麦、大麦及小黑麦花药预处理、胚状体和愈伤诱导、绿苗再生及白苗形成遗传控制方面进行了大量的研究。本文对这些研究进行综述,以期为相关研究提供参考。 相似文献
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Rice is a model system used for crop genomics studies. The completion of the rice genome draft sequences in 2002 not only accelerated functional genome studies, but also initiated a new era of resequencing rice genomes. Based on the reference genome in rice, next-generation sequencing (NGS) using the high-throughput sequencing system can efficiently accomplish whole genome resequencing of various genetic populations and diverse germplasm resources. Resequencing technology has been effectively utilized in evolutionary analysis, rice genomics and functional genomics studies. This technique is beneficial for both bridging the knowledge gap between genotype and phenotype and facilitating molecular breeding via gene design in rice. Here, we also discuss the limitation, application and future prospects of rice resequencing. 相似文献